Generation of a nucleosome-free promoter region in SV40 does not require T-antigen binding to site I.

The relationship between T-antigen interactions at Site I and the presence of a nucleosome-free promoter in SV40 chromatin was examined by analyzing chromatin from mutants defective for T-antigen interaction at site I (cs 1085, scs111, and tsA58) and their parental wild-type strains (776, SVS, and VA45-54, respectively). As judged by sensitivity to digestion with restriction endonucleases that recognize unique sequences within the promoter region (BglI, KpnI, and MspI), a nucleosome-free promoter was observed in a substantially larger proportion of chromosomes from the defective mutants than their wild-type parents. This result demonstrates that T-antigen binding to site I is not necessary for setting the early boundary of the nucleosome-free region, although it may function directly or indirectly in determining the proportion of chromosomes containing this feature.